Cyclodextrin can increase the solubility of poorly soluble drugs, but also decrease the permeability of poorly soluble drugs in inclusion complexes simultaneously, which partially or completely counteracts the contribution of improvement in solubility to the oral absorption of poorly soluble drugs. If a competing agent is added to the system to compete binding sites of cyclodextrins with drugs, drug permeability can be improved by increasing the concentration of free drugs in the inclusion complex system. In this paper, a rapid in vitro screening method for competing agents of cyclodextrin inclusion complex is proposed based on the principle that good drug permeability is in accord with good cell uptake. The equilibrium constants between drugs and hydroxypropyl-beta-cyclodextrin (HPCD) were determined by phase equilibrium solubility method. Cinnarizine (CN) with a high equilibrium constant was selected as a competing agent, coumarin 6 (C6) and 9-octadecyl berberine (BD) with smaller equilibrium constants were selected as model drugs. Both changes of solubility and uptake by Caco-2 and A549 cells of C6 and BD were investigated different concentrations of CN to the HPCD solution of C6 and BD. The results showed that the uptake of C6 and BD increased in a CN concentration-dependent manner, and the solubility of C6 and BD in HPCD solution decreased with the prolongation of equilibrium time. It might be due to increased free drug concentrations that resulted from the competition of CN for drug binding sites with HPCD. In our study, in vitro cell uptake method was firstly used to validate the ability of CN as a competing agent to increase drug permeability (cell uptake). This method can be used for preliminarily screening of competing agents for drug-cyclodextrin inclusion complexes.

BCS Ⅱ drugs are characterized by low solubility and high permeability. Improving their solubility is considered an important approach to improve its oral absorption. Recent strategies to increase the solubility of poorly-soluble drugs may unexpectedly result in greatly depressed permeability, ultimately leading to failure in improving oral absorption. Based on the mathematics of membrane permeability coefficient of a drug, the membrane/aqueous partition coefficient is dependent on the drug's solubility in the gastrointestinal milieu, suggesting a unique interplay between the solubility and permeability of the drug, and treating the one irrespectively of the other may be insufficient. When we focus on the increase of drug solubility and overlook the efficacy of drug permeability, the positive effect of increased solubility to drug oral absorption might be traded off by depressed permeability. To provide rational formulary designs, by optimizing excipients and evaluation, this review summarizes solubility- permeability interplay for different types of solubilizing techniques, such as cyclodextrin, surfactants-based vehicle, cosolvent, amorphous solid dispersions, other infectors such as P-gp transporters and new techniques for simultaneous evaluation of drug solubility and permeability.

OBJECTIVETo investigate the effects of lead exposure to rat placenta and pups during different gestation periods.

METHODSAll 108 Wistar rats (72 females, 36 males) were randomly divided into four groups. All rats were orally fed with 0.025% lead acetate during different gestation periods. Blood was obtained from the abdominal vena cava and the lead level in maternal blood was measured by means of atomic absorption spectrometry at the end of the pregnancy. The number of pups, their body weight, body length and tail length were measured. The effects of lead to rat placenta were observed by level of microscopy, optical microscopy and electronic microscopy.

RESULTSExperimental groups the blood lead level at the end of gestation were above 0.483 micromol/L. There were significant differences among, of pups, during different groups (P < 0.01). Among them the drinking lead group of whole distant was the lowest in placenta weight [(0.31 +/- 0.13) g] body weight of pups [(2.08 +/- 0.88) g] length and tail length of pups [(2.37 +/- 0.32) cm, (0.98 +/- 0.09) cm]. There were significantly differences between the experimental groups and controls. Maternal blood lead level was negatively related to placenta weight (r = 0.652, P < 0.01), and had no relation with the body weight of pups (r = -0.107, P = 0.46). In the experimental groups of lead poisoned rats, the placenta showed focus necrosis in the deciduas, and increased the trophoblastic giant cells and light staining cells in the trophospongium. Trophoblast in the labyrinth and trophospongium showed degeneration; fibrin deposition around the villi was increased. Microvilli around the trophoblast were shorter and less, mitochondrion was swollen and decreased in number, rough endoplasmic reticulum was distended and ribosomal number on membrane decreased.

CONCLUSIONLead exposure during different gestation periods should have a traumatic effect on the trophoblast, leading to interference of nutrition and oxygen exchange. Furthermore, the blood supply to the placenta and nutrition and oxygen exchange between mother and pups were also interfered, leading to reduction of placenta weight and retardation of development of pups.

Animals ; Environmental Exposure ; adverse effects ; Female ; Lead ; toxicity ; Male ; Organ Size ; drug effects ; Placenta ; drug effects ; Pregnancy ; Rats ; Rats, Wistar

BACKGROUNDThe diagnosis of Parkinson's disease is presently based on non-specific symptoms. However, radionuclide dopamine transporters imaging can provide specific diagnostic tool for Parkinson's disease. This study was designed to investigate the effects of imaging of dopamine transporters with 99mTc-TRODAT-1 in early diagnosis or differential diagnosis of Parkinson's disease.

METHODSNine normal monkeys were used to establish N-methyl-4-phenyl-1, 2, 3, 6-tetra-hydropyridine (MPTP) hemi-Parkinsonian animal models, and they were subjected to imaging. Twenty-nine patients with Parkinson's disease, 12 age-matched healthy volunteers, and 18 age-matched patients with Parkinson's syndrome were investigated. Single photon emission computer tomography (SPECT) was performed 3 hours after intravenous injection of 740 MBq 99mTc-TRODAT-1. Striatum specific uptake of 99mTc-TRODAT-1 was calculated according to the ratio of striatum (ST) to cerebellum (CB) in dopamine transporters uptake.

RESULTSIn normal monkeys, bilateral ratio of ST/CB was 2.34 +/- 0.41. After the injection of MPTP, uptake rate of 99mTc-TRODAT-1 at damaged region was much lower than that at the contralateral region, resulting in a significant difference in the ratio of ST/CB (right: ST/CB = 1.73 +/- 0.35; left: ST/CB = 1.90 +/- 0.30), especially in hemi-Parkinsonian model monkeys (right: ST/CB = 1.29 +/- 0.17; left: ST/CB = 1.80 +/- 0.33). The ratios of ST/CB were 1.57 +/- 0.17 and 1.61 +/- 0.14 for the right and left respectively in the healthy volunteers, 1.04 +/- 0.29 and 1.06 +/- 0.30 in the age-matched patients with Parkinson's disease, and 1.56 +/- 0.17 and 1.59 +/- 0.18 in the age-matched patients with Parkinson's disease syndrome. A significant difference was noted between group of Parkinson's disease, normal controls and Parkinson's disease syndrome.

CONCLUSIONThe results suggest that 99mTc-TRODAT-1 dopamine transporters SPECT has clinical application value in early diagnosis or differential diagnosis of Parkinson's disease.

Adult ; Aged ; Animals ; Dopamine Plasma Membrane Transport Proteins ; Female ; Haplorhini ; Humans ; Male ; Membrane Glycoproteins ; analysis ; Membrane Transport Proteins ; analysis ; Middle Aged ; Nerve Tissue Proteins ; analysis ; Organotechnetium Compounds ; Parkinson Disease ; diagnosis ; Radiopharmaceuticals ; Tomography, Emission-Computed, Single-Photon ; Tropanes

Objective:To study therapeutic effect of trimetazidine on patients with angina pectoris after percutaneous coronary intervention(PCI)and its influence on endothelial function and exercise tolerance.Methods:A total of 108 patients with angina pectoris after primary PCI in our hospital were selected,randomly and equally divided into routine treatment group and trimetazidine group(received trimetazidine therapy since second month after PCI based on routine treatment).Therapeutic effect after treatment,endothelial function and exercise tolerance before and after treatment were compared between two groups.Results:Total effective rate of trimetazidine group was signifi-cantly higher than that of routine treatment group(98.15% vs.83.33%,P=0.008).Compared with before treat-ment,there were significant improvements in angina pectoris onset condition and cardiac function,significant rise in endothelial function and exercise tolerance in two groups after treatment,P<0.05 or <0.01;compared with rou-tine treatment group after treatment,there were significant reductions in mean onset times[(2.91 ± 1.25)times/month vs.(1.72 ± 1.04)times/month]and duration[(2.06 ± 1.08)h vs.(1.24 ± 0.97)h]of angina pectoris,ni-troglycerin dose[(1.21 ± 0.12)g vs.(0.84 ± 0.11)g],depression magnitude[(0.16 ± 0.05)mV vs.(0.13 ± 0.05)mV]and time[(18.64 ± 6.41)min vs.(6.74 ± 4.82)min]of ST segment,T wave inversion magnitude [(0.18 ± 0.04)mV vs.(0.09 ± 0.05)mV]and endothelin-1 level[(50.44 ± 7.43)ng/L vs.(41.08 ± 6.02)ng/L],and significant rise in left ventricular ejection fraction[(54.18 ± 8.25)% vs.(57.82 ± 7.73)%],6min walking distance[(512.74 ± 86.66)m vs.(551.24 ± 84.69)m],fore brachial artery endothelium dependent diastolic-sys-tolic function[(10.08 ± 3.1)% vs.(11.79 ± 3.24)%]and nitric oxide level[(83.14 ± 5.15)μmol/L vs.(116.22 ± 6.54)μmol/L]in trimetazidine group,P<0.05 or <0.01. Conclusion:Trimetazidine possesses good treatment effect on angina pectoris after PCI,and it can effectively improve endothelial function and exercise tolerance,which is worth extending.

OBJECTIVETo clone 1b type of HCV NS3-4b Gene and express in HEK 293 cells, lay the foundation for further study of the HCV NS3-4b recombinant adeno-associated virus vaccine and its dendritic cell vaccine.

METHODSHCV 1b patients' serum was collected, and full length NS3-4b segment was amplified by RT-PCR and cloned into adeno-associated virus' expression vector pAAV. CMV. EGFP in order to express in HEK 293 cells. At last, it was validated whether express or not by Western Blot.

RESULTSThe 1b type gene NS3-4b were amplified and consistent to the expected size (2838 bp), the recombinant plasmid has been confirmed its successful restructured by double enzyme and sequencing, at last, Western Blot map can see objective protein expression after it transfect HEK 293 cells.

CONCLUSIONThe adeno-associsted virus recombination HCV NS3-4b plasmid have successfully constructed and it can express in eukaryotic cells.

Dependovirus ; genetics ; Genetic Vectors ; HEK293 Cells ; Hepacivirus ; genetics ; Humans ; Plasmids ; Vaccines, Synthetic ; immunology ; Viral Nonstructural Proteins ; genetics ; Viral Vaccines ; immunology

This study was aimed to determine the hepatic and small intestinal distribution of active lignans in rats after treated with Wuzhi tablet (WZ, Schisandra sphenanthera extract) by LC-MS/MS method. Male Sprague-Dawley rats were sacrificed at 0.25, 1.5, 4, 6, 10, 24 h after an oral administration of WZ, and then hepatic and small intestinal samples were collected for analysis. The results showed that concentrations of lignans in liver and small intestine of rats were decreased with WZ pretreated time. The concentrations of all lignans in rat liver and small intestine at 0.25 h were the highest after a single oral administration. All lignans was undetectable in all tissues 24 h after oral dosing, suggesting lignans of WZ were eliminated rapidly in rats. The concentrations of schisandrin A, schisandrol B and schisantherin A in small intestine were much higher than those in the liver, suggesting the effect of WZ on the intestinal metabolism enzyme might be more potent than that on the liver. In short, the current results suggest that lignans of WZ were not accumulated in rat liver and small intestine. The concentrations of lignans of WZ in small intestine were much higher than those in liver.

ObjectiveTo summarize and analyze the clinical characteristics， diagnosis process， treatment process， and obstetric outcomes of pregnant women with Cushing's syndrome， helping to optimize pregnancy management. MethodsA retrospective study was conducted on 8 pregnant women with Cushing’s syndrome who were hospitalized in the First Affiliated Hospital， Sun Yat-sen University between January 2006 and August 2022. The clinical characteristics， management and obstetric outcomes were recorded. ResultsPreeclampsia was detected in 4 cases，pre-gestational diabetes mellitus in 2 cases， gestational diabetes mellitus in 5 cases， and hypokalemia in all 8 cases. Elevated serum cortisol， disappearance of day-night rhythm of cortisol， increased 24-hour urine cortisol and decrease in serum ACTH were found in 8 cases by laboratory examination. Furthermore， adrenal adenoma was detected in all 8 cases by ultrasonography or Magnetic Resonance Imaging. Three cases underwent laparoscopic adrenalectomy in the second trimester and 4 cases received surgery after delivery. The diagnosis of adrenal cortical adenoma was confirmed by pathological report. Six cases had preterm birth， while one patient delivered after 37 weeks of gestation and one patient suffered from spontaneous abortion. Among 7 cases of live birth， 6 patients underwent cesarean section and 1 patient had vaginal delivery. Of all newborns， 3 had low birth weight. One case had a birth defect. Four infants were transferred to the neonatal intensive care unit， and two infants died. One child was diagnosed with nephrotic syndrome at 2 years of age. ConclusionsCushing's syndrome is rare and high risk during pregnancy. It requires multidisciplinary diagnosis， treatment， and long-term follow-up. Drug therapy carries a risk of progression and requires intensive care during pregnancy， postpartum follow-up， and specialist treatment.

This study was purposed to compare the anti-leukemic effects of E.coli-L-Asp and Erwinia-L-Asp in vitro, and to investigate their mechanism. The cell apoptosis and proliferation inhibition rate were measured by CCK-8 kit, and IC50 of two drugs was calculated by using SPSS software. Pro-apoptosis effect of E.coli-L-Asp and Erwinia-L-Asp on REH and Jurkat cell lines was also determined by flow cytometry with Annexin V/PI double staining. Concentration changes of 4 amino acids (Asn, Aspa, Gln, and Glu) before and after medication were detected by using high pressure liquid chromatography (HPLC) assay. The results showed that both REH and Jurkat cell lines were sensitive to L-Asp drugs from two different strains, and E.coli-L-Asp and Erwinia-L-Asp displayed the inhibition effect on the proliferation of Jurkat and REH cell lines in dose-dependent and time-dependent manners. The inhibition cell of proliferation and cell apoptosis in Erwinia-L-Asp group were higher than those in E.coli-L-Asp group after 24 hours (P < 0.05) . However, after treatment of REN and Jurkat cells with 2 kind of L-Asp for 48 hours, the inhibition of cell proliferation and apoptosis rates were not significantly different between the 2 L-Asp drugs (P > 0.05). The Asn in medium could be depleted by two different L-Asp drugs with low concentration. Both the two L-Asp drugs had the same capability to deplete the Asn surrounding leukemia cells (P > 0.05). The Gln in medium could be depleted by two L-Asp drugs with high concentration. The hydrolysis effect of Erwinia-L-Asp on Gln was stronger than that of E.coli-L-Asp (P < 0.05). It is concluded that in a certain range of concentrations, E.coli-L-Asp and Erwinia-L-Asp exert anti-leukemia effect in dose-dependent manner. Depletion of Gln and Asn in surrounding environment and induction of cell apoptosis are two potential mechanisms, by which leukemia cells can be killed. Erwinia-L-Asp may be chosen as the first-line drug to treat childhood ALL for its fast action and stronger hydrolysis effect on Gln.
Apoptosis ; drug effects ; Asparaginase ; pharmacology ; Cell Proliferation ; drug effects ; Flow Cytometry ; Humans ; Jurkat Cells

OBJECTIVETo study the feasibility of vector-mediated RNA interference for HER-2-positive breast cancer therapy.

METHODSA plasmid vector capable of mediating HER-2 RNA interference was constructed, and HER-2-positive breast cancer cell line SKBR-3 was transfected with this constructed vector. The expression of HER-2 mRNA and protein was analyzed by RT-PCR and Western blotting, and the growth and apoptosis of SKBR-3 cells was analyzed after transfection.

RESULTSThe expressions of HER-2 mRNA and HER-2 protein was downregulated in response to vector-mediated HER-2 RNA interference, which also resulted in tumor cell growth inhibition and increased number apoptotic cells.

CONCLUSIONHER-2 is a good target for RNA interference and RNA interference targeting HER-2 can lead to HER-2 breast cancer cell apoptosis and growth inhibition.

Apoptosis ; Blotting, Western ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Female ; Genetic Vectors ; Humans ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Receptor, ErbB-2 ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection